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两种人结肠癌细胞生长和分化过程中胰蛋白酶原I的表达

Pancreatic trypsinogen I expression during cell growth and differentiation of two human colon carcinoma cells.

作者信息

Bernard-Perrone F, Carrere J, Renaud W, Moriscot C, Thoreux K, Bernard P, Servin A, Balas D, Senegas-Balas F

机构信息

Groupe de Recherche sur la Trophicité et le Vieillissement, Faculté de Médecine, Nice, France.

出版信息

Am J Physiol. 1998 Jun;274(6):G1077-86. doi: 10.1152/ajpgi.1998.274.6.G1077.

Abstract

Pancreatic trypsin has been found to induce tight junction or dome formation in some colon cancer cell lines (HT-29, Caco-2), and a tumor-associated trypsinogen, trypsinogen type II, has been isolated from another colon cancer cell line (COLO 205). We have tried to determine if trypsinogen is present and how its expression varies during cell culture in HT-29 Glc+/- and Caco-2 cells, which exhibit enterocytic differentiation, and in HT-29 Glc+ cells, which never differentiate. Trypsinogen mRNA presence and expression were demonstrated in these cells by mRNA hybridization, RT-PCR, cytoimmunofluorescence, Western immunoblot analysis, and gel filtration. Trypsinogen was found to be trypsinogen type I and was mainly in zymogen form in culture media. Differentiating cells exhibited variations in trypsinogen I expression, but cells that remained undifferentiated did not. In the differentiated cells, a high and transient peak in trypsinogen I expression was observed during the first steps of differentiation.

摘要

已发现胰腺胰蛋白酶可在某些结肠癌细胞系(HT - 29、Caco - 2)中诱导紧密连接或穹顶形成,并且已从另一种结肠癌细胞系(COLO 205)中分离出一种肿瘤相关胰蛋白酶原,即II型胰蛋白酶原。我们试图确定在表现出肠细胞分化的HT - 29 Glc+/-和Caco - 2细胞以及从不分化的HT - 29 Glc+细胞的细胞培养过程中是否存在胰蛋白酶原及其表达如何变化。通过mRNA杂交、逆转录聚合酶链反应(RT - PCR)、细胞免疫荧光、蛋白质免疫印迹分析和凝胶过滤证明了这些细胞中存在胰蛋白酶原mRNA并表达。发现胰蛋白酶原为I型胰蛋白酶原,且在培养基中主要以酶原形式存在。分化细胞的I型胰蛋白酶原表达存在差异,但未分化细胞则没有。在分化细胞中,在分化的第一步观察到I型胰蛋白酶原表达出现高且短暂的峰值。

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